{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T10:16:58Z","timestamp":1782728218034,"version":"3.54.5"},"reference-count":25,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2021,6,25]],"date-time":"2021-06-25T00:00:00Z","timestamp":1624579200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>A universal influenza candidate vaccine that targets multiple conserved influenza virus epitopes from hemagglutinin (HA), neuraminidase (NA) and matrix (M2e) proteins was combined with the potent Army liposomal adjuvant (ALFQ) to promote induction of broad immunity to seasonal and pandemic influenza strains. The unconjugated and CRM-conjugated composite peptides formulated with ALFQ were highly immunogenic and induced both humoral and cellular immune responses in mice. Broadly reactive serum antibodies were induced across various IgG isotypes. Mice immunized with the unconjugated composite peptide developed antibody responses earlier than mice immunized with conjugated peptides, and the IgG antibodies were broadly reactive and neutralizing across Groups 1 and 2 influenza viruses. Multi-epitope unconjugated influenza composite peptides formulated with ALFQ provide a novel strategy for the development of a universal influenza vaccine. These synthetic peptide vaccines avoid the pitfalls of egg-produced influenza vaccines and production can be scaled up rapidly and economically.<\/jats:p>","DOI":"10.3390\/vaccines9070698","type":"journal-article","created":{"date-parts":[[2021,6,25]],"date-time":"2021-06-25T11:07:40Z","timestamp":1624619260000},"page":"698","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Conserved Influenza Hemagglutinin, Neuraminidase and Matrix Peptides Adjuvanted with ALFQ Induce Broadly Neutralizing Antibodies"],"prefix":"10.3390","volume":"9","author":[{"given":"Clara J.","family":"Sei","sequence":"first","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, Gaithersburg, MD 20878, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mangala","family":"Rao","sequence":"additional","affiliation":[{"name":"U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard F.","family":"Schuman","sequence":"additional","affiliation":[{"name":"Antibody and Immunoassay Consultants, Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke T.","family":"Daum","sequence":"additional","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, San Antonio, TX 78209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2074-2373","authenticated-orcid":false,"given":"Gary R.","family":"Matyas","sequence":"additional","affiliation":[{"name":"U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nimisha","family":"Rikhi","sequence":"additional","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, Gaithersburg, MD 20878, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin","family":"Muema","sequence":"additional","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, Gaithersburg, MD 20878, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0000-7734","authenticated-orcid":false,"given":"Alexander","family":"Anderson","sequence":"additional","affiliation":[{"name":"U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"},{"name":"Oak Ridge Institute of Science and Education, Oak Ridge, TN 37831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ousman","family":"Jobe","sequence":"additional","affiliation":[{"name":"U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"},{"name":"Henry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kellie A.","family":"Kroscher","sequence":"additional","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, Gaithersburg, MD 20878, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carl R.","family":"Alving","sequence":"additional","affiliation":[{"name":"U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerald W.","family":"Fischer","sequence":"additional","affiliation":[{"name":"Longhorn Vaccines and Diagnostics, Gaithersburg, MD 20878, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,25]]},"reference":[{"key":"ref_1","unstructured":"World Health Organisation (2021, June 11). Influenza (Seasonal). Available online: http:\/\/www.who.int\/en\/news-room\/fact-sheets\/detail\/influenza-(seasonal)."},{"key":"ref_2","unstructured":"World Health Organisation (2020, June 11). Cumulative Number of Confirmed Human Cases for Avian Influenza A(H5N1) Reported to WHO 2003\u20132020. Available online: https:\/\/www.who.int\/influenza\/human_animal_interface\/2020_01_20_tableH5N1.pdf?ua=1."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1002\/jmv.10143","article-title":"Divergent evolution of hemagglutinin and neuraminidase genes in recent influenza A: H3N2 viruses isolated in Canada","volume":"67","author":"Abed","year":"2002","journal-title":"J. Med. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1111\/j.1750-2659.2011.00304.x","article-title":"Influenza neuraminidase","volume":"6","author":"Air","year":"2012","journal-title":"Influ. Other Respir. Viruses"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.cell.2018.03.030","article-title":"Influenza Infection in Humans Induces Broadly Cross-Reactive and Protective Neuraminidase-Reactive Antibodies","volume":"173","author":"Chen","year":"2018","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2067","DOI":"10.1093\/cid\/ciy818","article-title":"Neutralizing Antibody Responses to Homologous and Heterologous H1 and H3 Influenza A Strains After Vaccination with Inactivated Trivalent Influenza Vaccine Vary with Age and Prior-year Vaccination","volume":"68","author":"Wang","year":"2018","journal-title":"Clin. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"S75","DOI":"10.1093\/infdis\/jiz017","article-title":"Neuraminidase, the Forgotten Surface Antigen, Emerges as an Influenza Vaccine Target for Broadened Protection","volume":"219","author":"Eichelberger","year":"2019","journal-title":"J. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"105","DOI":"10.3390\/vaccines3010105","article-title":"M2e-Based Universal Influenza A Vaccines","volume":"3","author":"Deng","year":"2015","journal-title":"Vaccines"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1038\/13484","article-title":"A Universal Influenza Vaccine Based on the Extracellular Domain of the M2e Protein","volume":"5","author":"Neirynck","year":"1999","journal-title":"Nat. Med."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Tsilibary, E.-P., Charonis, S.A., and Georgopoulos, A.P. (2021). Vaccines for Influenza. Vaccines, 9.","DOI":"10.3390\/vaccines9010047"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1126\/science.1171491","article-title":"Antibody Recognition of a Highly Conserved Influenza Virus Epitope","volume":"324","author":"Ekiert","year":"2009","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-28706-1","article-title":"The influenza virus hemagglutinin head evolves faster than the stalk domain","volume":"8","author":"Kirkpatrick","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3700","DOI":"10.1128\/JVI.02576-14","article-title":"Structure of the Extracellular Domain of Matrix Protein 2 of Influenza A Virus in Complex with a Protective Monoclonal Antibody","volume":"89","author":"Cho","year":"2015","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"McMillan, C.L., Young, P.R., Watterson, D., and Chappell, K.J. (2021). The Next Generation of Influenza Vaccines: Towards a Universal Solution. Vaccines, 9.","DOI":"10.3390\/vaccines9010026"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1111\/j.1600-065X.1995.tb00075.x","article-title":"Liposomes as Carriers of Peptide Antigens: Induction of Antibodies and Cytotoxic T Lymphocytes to Conjugated and Unconjugated Peptides","volume":"145","author":"Alving","year":"1995","journal-title":"Immunol. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.1016\/0264-410X(94)00058-U","article-title":"Antibody and cytotoxic T-lymphocyte responses to a single liposome-associated peptide antigen","volume":"13","author":"White","year":"1995","journal-title":"Vaccine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2804","DOI":"10.1016\/j.vaccine.2013.04.027","article-title":"Lip-osomes containing monophosphoryl lipid A: A potent adjuvant system for inducing antibodies to heroin hapten analogs","volume":"31","author":"Matyas","year":"2013","journal-title":"Vaccine"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1021\/acs.jmedchem.7b01427","article-title":"A Stable Heroin Analogue That Can Serve as a Vaccine Hapten to Induce Antibodies That Block the Effects of Heroin and Its Metabolites in Rodents and That Cross-React Immunologically with Related Drugs of Abuse","volume":"11","author":"Sulima","year":"2018","journal-title":"J. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41541-017-0013-9","article-title":"Heroin-HIV-1 (H2) vaccine: Induction of dual immunologic effects with a heroin hapten-conjugate and an HIV-1 envelope V2 peptide with liposomal lipid A as an adjuvant","volume":"2","author":"Torres","year":"2017","journal-title":"NPJ Vaccines"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1586\/erv.12.35","article-title":"Liposomes containing lipid A: An effective, safe, generic ad-juvant system for synthetic vaccines","volume":"11","author":"Alving","year":"2012","journal-title":"Expert Rev. Vaccines"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5578","DOI":"10.1016\/j.vaccine.2015.09.001","article-title":"Differential immune responses to HIV-1 envelope protein induced by liposomal adjuvant formulations containing monophosphoryl lipid A with or without QS21","volume":"33","author":"Beck","year":"2015","journal-title":"Vaccine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/14760584.2020.1745636","article-title":"Army Liposome Formulation (ALF) family of vaccine adjuvants","volume":"19","author":"Alving","year":"2020","journal-title":"Expert Rev. Vaccines"},{"key":"ref_23","unstructured":"Hermanson, G.T. (2013). Bioconjugate Techniques, Elsevier."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1039\/C5SC03892H","article-title":"Peptide-based synthetic vaccines","volume":"7","author":"Skwarczynski","year":"2016","journal-title":"Chem. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.coviro.2020.02.003","article-title":"Tracking progress in universal influenza vaccine development","volume":"40","author":"Ostrowsky","year":"2020","journal-title":"Curr. Opin. Virol."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/7\/698\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:24:08Z","timestamp":1760163848000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/7\/698"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,25]]},"references-count":25,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2021,7]]}},"alternative-id":["vaccines9070698"],"URL":"https:\/\/doi.org\/10.3390\/vaccines9070698","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,25]]}}}